FIG 6.

MxB disrupts interaction of CypA with HCV protein NS5A. (A) Effects of MxB and its mutants on interaction of NS5A with CypA. MxB-Myc, NS5A-Flag, and CypA-HA were coexpressed. Immunoprecipitation was performed to pull down CypA-HA by use of anti-HA antibody. The level of coprecipitated NS5A-Flag was determined by Western blotting (IB). (B) Protein band intensities in the Western blots in panel A were determined by use of NIH ImageJ software. Data are normalized to the control group, with the control value arbitrarily set to 1. (C) Colocalization of NS5A and CypA is disrupted by MxB. Huh7.5.1 cells were cotransfected with NS5A-Flag, CypA-HA, and MxB-Myc or its mutants. Cells were fixed and stained with anti-Flag (red), anti-HA (green), and anti-Myc (blue) antibodies. Representative images are shown. Bars, 5 μm. (D) Colocalization of CypA (green) and NS5A (red) was assessed for randomly selected cells (n > 30) by use of Image-Pro Plus 7.0C software. Data are representative of at least three independent experiments, and values are expressed as means ± SD.